Linked Life Data

16689533

Source:http://linkedlifedata.com/resource/pubmed/id/16689533

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
10
pubmed:dateCreated
2006-5-12
pubmed:abstractText
Nucleic acid assay from a complex biological milieu is attractive but currently difficult and far from routine. In this study, DNA hybridization from serum dilutions into mixed DNA/mercaptoundecanol (MCU) adlayers on gold was monitored by surface plasmon resonance (SPR). Immobilized DNA probe and hybridized target densities on these surfaces were quantified using 32P-radiometric assays as a function of MCU diluent exposure. SPR surface capture results correlated with radiometric analysis for hybridization performance, demonstrating a maximum DNA hybridization on DNA/MCU mixed adlayers. The maximum target surface capture produced by MCU addition to the DNA probe layer correlates with structural and conformational data on identical mixed DNA/MCU adlayers on gold derived from XPS, NEXAFS, and fluorescence intensity measurements reported in a related study (Lee, C.-Y.; Gong, P.; Harbers, G. M.; Grainger, D. W.; Castner, D. G.; Gamble, L. J. Anal. Chem. 2006, 78, 3316-3325.). MCU addition into the DNA adlayer on gold also improved surface resistance to both nonspecific DNA and serum protein adsorption. Target DNA hybridization from serum dilutions was monitored with SPR on the optimally mixed DNA/MCU adlayers. Both hybridization kinetics and efficiency were strongly affected by nonspecific protein adsorption from a complex milieu even at a minimal serum concentration (e.g., 1%). No target hybridization was detected in SPR assays from serum concentrations above 30%, indicating nonspecific protein adsorption interference of DNA capture and hybridization from complex milieu. Removal of nonsignal proteins from nucleic acid targets prior to assay represents a significant issue for direct sample-to-assay nucleic acid diagnostics from food, blood, tissue, PCR mixtures, and many other biologically complex sample formats.
pubmed:grant
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-10920027, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-11283596, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-12443474, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-12708875, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-1420304, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-14961746, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-15568868, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-15734557, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-15743088, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-15896061, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-16390092, http://linkedlifedata.com/resource/pubmed/commentcorrection/16689533-16689533
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0003-2700
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
78
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3326-34
pubmed:dateRevised
2011-5-4
pubmed:meshHeading
pubmed:year
2006
pubmed:articleTitle
Hybridization behavior of mixed DNA/alkylthiol monolayers on gold: characterization by surface plasmon resonance and 32P radiometric assay.
pubmed:affiliation
Department of Chemistry, Colorado State University, Fort Collins, CO 80523-1872, USA.
pubmed:publicationType
Journal Article, Research Support, N.I.H., Extramural